LOCUS       V045263                 9387 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045263
VERSION     V045263
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             1..634
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     misc_feature    681..806
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1303..1536
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1721..1765
                     /codon_start=1
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /product="antigenic peptide corresponding to amino acids
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
                     et al., 2013)"
                     /transl_table=1
                     /translation="KNEQELLELDKWASL"
                     /label="gp41 peptide"
     misc_feature    2028..2143
                     /note="central polypurine tract and central termination
                     sequence of HIV-1 (lacking the first T)"
                     /label="cPPT/CTS"
     enhancer        2201..2504
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2505..2708
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2827..3411
                     /label="PYCARD(NM_013258)"
                     /note="PYCARD(NM_013258)"
                     /gene="PYCARD"
     CDS             3442..4158
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric green fluorescent protein, also known
                     as mTagGFP"
                     /transl_table=1
                     /translation="MSGGEELFAGIVPVLIELDGDVHGHKFSVRGEGEGDADYGKLEIK
                     FICTTGKLPVPWPTLVTTLCYGIQCFARYPEHMKMNDFFKSAMPEGYIQERTIQFQDDG
                     KYKTRGEVKFEGDTLVNRIELKGKDFKEDGNILGHKLEYSFNSHNVYIRPDKANNGLEA
                     NFKTRHNIEGGGVQLADHYQTNVPLGDGPVLIPINHYLSTQTKISKDRNEARDHMVLLE
                     SFSACCHTHGMDELYR*"
                     /label="TagGFP2"
     promoter        4175..4674
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4695..5294
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     misc_feature    5308..5896
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6103..6736
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6865..6881)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6889..6905
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(6913..6943)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6958..6979
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7267..7855)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8026..8886)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(8887..8991)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9039..9173
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa
     2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     2761 tccatagaag acaccgactc tactagagga tcgctagcgc taccggactc agatctcgag
     2821 gccaccatgg ggcgcgcgcg cgacgccatc ctggatgcgc tggagaacct gaccgccgag
     2881 gagctcaaga agttcaagct gaagctgctg tcggtgccgc tgcgcgaggg ctacgggcgc
     2941 atcccgcggg gcgcgctgct gtccatggac gccttggacc tcaccgacaa gctggtcagc
     3001 ttctacctgg agacctacgg cgccgagctc accgctaacg tgctgcgcga catgggcctg
     3061 caggagatgg ccgggcagct gcaggcggcc acgcaccagg gctctggagc cgcgccagct
     3121 gggatccagg cccctcctca gtcggcagcc aagccaggcc tgcactttat agaccagcac
     3181 cgggctgcgc ttatcgcgag ggtcacaaac gttgagtggc tgctggatgc tctgtacggg
     3241 aaggtcctga cggatgagca gtaccaggca gtgcgggccg agcccaccaa cccaagcaag
     3301 atgcggaagc tcttcagttt cacaccagcc tggaactgga cctgcaagga cttgctcctc
     3361 caggccctaa gggagtccca gtcctacctg gtggaggacc tggagcggag cggaggagga
     3421 ggatcaggag gaggaggatc catgagcggg ggcgaggagc tgttcgccgg catcgtgccc
     3481 gtgctgatcg agctggacgg cgacgtgcac ggccacaagt tcagcgtgcg cggcgagggc
     3541 gagggcgacg ccgactacgg caagctggag atcaagttca tctgcaccac cggcaagctg
     3601 cccgtgccct ggcccaccct ggtgaccacc ctctgctacg gcatccagtg cttcgcccgc
     3661 taccccgagc acatgaagat gaacgacttc ttcaagagcg ccatgcccga gggctacatc
     3721 caggagcgca ccatccagtt ccaggacgac ggcaagtaca agacccgcgg cgaggtgaag
     3781 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcaaggactt caaggaggac
     3841 ggcaacatcc tgggccacaa gctggagtac agcttcaaca gccacaacgt gtacatccgc
     3901 cccgacaagg ccaacaacgg cctggaggct aacttcaaga cccgccacaa catcgagggc
     3961 ggcggcgtgc agctggccga ccactaccag accaacgtgc ccctgggcga cggccccgtg
     4021 ctgatcccca tcaaccacta cctgagcact cagaccaaga tcagcaagga ccgcaacgag
     4081 gcccgcgacc acatggtgct cctggagtcc ttcagcgcct gctgccacac ccacggcatg
     4141 gacgagctgt acaggtaatc tagataattc taccgggtag gggaggcgct tttcccaagg
     4201 cagtctggag catgcgcttt agcagccccg ctgggcactt ggcgctacac aagtggcctc
     4261 tggcctcgca cacattccac atccaccggt aggcgccaac cggctccgtt ctttggtggc
     4321 cccttcgcgc caccttctac tcctccccta gtcaggaagt tcccccccgc cccgcagctc
     4381 gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca ctagtctcgt gcagatggac
     4441 agcaccgctg agcaatggaa gcgggtaggc ctttggggca gcggccaata gcagctttgc
     4501 tccttcgctt tctgggctca gaggctggga aggggtgggt ccgggggcgg gctcaggggc
     4561 gggctcaggg gcggggcggg cgcccgaagg tcctccggag gcccggcatt ctgcacgctt
     4621 caaaagcgca cgtctgccgc gctgttctcc tcttcctcat ctccgggcct ttcgacctgc
     4681 agcccaagct taccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg
     4741 tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca
     4801 ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc
     4861 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct
     4921 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg
     4981 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc
     5041 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg
     5101 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg
     5161 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct
     5221 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca
     5281 agcccggtgc ctgaccgcgt ctggaacaat caacctctgg attacaaaat ttgtgaaaga
     5341 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     5401 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     5461 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     5521 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     5581 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     5641 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     5701 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     5761 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     5821 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     5881 tgggccgcct ccccgcctgg aattaattct gcagtcgaga cctagaaaaa catggagcaa
     5941 tcacaagtag caatacagca gctaccaatg ctgattgtgc ctggctagaa gcacaagagg
     6001 aggaggaggt gggttttcca gtcacacctc aggtaccttt aagaccaatg acttacaagg
     6061 cagctgtaga tcttagccac tttttaaaag aaaagagggg actggaaggg ctaattcact
     6121 cccaacgaag acaagatatc cttgatctgt ggatctacca cacacaaggc tacttccctg
     6181 attagcagaa ctacacacca gggccagggg tcagatatcc actgaccttt ggatggtgct
     6241 acaagctagt accagttgag ccagataagg tagaagaggc caataaagga gagaacacca
     6301 gcttgttaca ccctgtgagc ctgcatggga tggatgaccc ggagagagaa gtgttagagt
     6361 ggaggtttga cagccgccta gcatttcatc acgtggcccg agagctgcat ccggagtact
     6421 tcaagaactg ctgatatcga gcttgctaca agggactttc cgctggggac tttccaggga
     6481 ggcgtggcct gggcgggact ggggagtggc gagccctcag atcctgcata taagcagctg
     6541 ctttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     6601 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     6661 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     6721 tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca
     6781 aagaaatgaa tatcagagag tgagaggcct tgacattgct agcgttttac cgtcgacctc
     6841 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     6901 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     6961 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     7021 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     7081 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     7141 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     7201 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     7261 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     7321 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     7381 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     7441 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     7501 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     7561 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     7621 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     7681 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     7741 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     7801 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7861 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7921 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7981 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     8041 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     8101 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     8161 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     8221 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     8281 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     8341 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     8401 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     8461 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     8521 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     8581 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     8641 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     8701 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     8761 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     8821 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8881 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8941 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     9001 ccgaaaagtg ccacctgacg tcgacggatc gggagatcaa cttgtttatt gcagcttata
     9061 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     9121 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcaactgga
     9181 taactcaagc taaccaaaat catcccaaac ttcccacccc ataccctatt accactgcca
     9241 attacctgtg gtttcattta ctctaaacct gtgattcctc tgaattattt tcattttaaa
     9301 gaaattgtat ttgttaaata tgtactacaa acttagtagt ttttaaagaa attgtatttg
     9361 ttaaatatgt actacaaact tagtagt
//
